| Co-Investigator(Kenkyū-buntansha) |
SHINDO Heisaburo Tokyo Pharmaceutical College, Assistant Professor, 薬学部, 助教授 (80138966)
SHIRAKIHARA Yasuo National Institute of Genetics, Assistant Professor, 遺伝情報研究センター, 助教授 (20150287)
HAKOSHIMA Toshio Nara Advanced Institute of Science and Technology, Professor, バイオサイセンス研究科, 教授 (00164773)
YAMAZAKI Toshio Osaka University, Assistant Professor, 蛋白質研究所, 助手 (60273710)
ISHII Shunsuke RIKEN Life-science Tsukuba Center, Researcher, 分子遺伝学研究室, 主任研究員 (00124785)
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| Research Abstract |
In order to analyze the specific DNA-recognition of DNA-binding proteins, we have determined three dimensional structures of many DNA-binding domains in their DNA bound states, for example, a protooncogene product, c-Myb, a yeast phosphate metabolic transcription factor, Pho4, a E. coli replication termination factor, Tus, a thymine dimer repair enzyme, T4 endonuclease V, and an interferon responsive transcription factor, IRF2 by using X-ray and NMR methods. In the sequence recognizing proteins, an α helix is located in the major groove of DNA and several amino acids from the helix are interacting with specific bases holding the Watson/Crick type base pairs. While, on the other hand, T4 endonuclease V, which recognizes a damaged DNA, disrupts a Watson/Crick type base pair and a base is flipped out. In the case of c-Myb DNA-binding domain, the dynamic structures of its free and DNA-bound states have been investigated by using NMR ; in the free state a flexible linker connects two subdomains and each subdomain is a structurally independent unit, while, in the DNA-bound state the two subdomains are fixed in the major groove of DNA and recognize a specific base sequence cooperatively. In addition, we have determined three dimensional structures of several DNA-binding proteins including RuvC, RNA polymerase subunit α , purine repressor PurR, OmpR, PhoB, telomere repeat binding factor hTRF1, a general transcription factor TFIIE in their DNA-free states and their DNA-binding modes have been discussed.
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